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. 2013 May 23;199(3):749–757. doi: 10.1111/nph.12304

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Copyright © 2013 New Phytologist Trust

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Fig. 3 — UV-crosslink of the E. coli-expressed GST and GST-NbGSTU4 with Bamboo mosaic virus (BaMV) RNAs. (a) Recombinant glutathione S-transferase (GST) and GST-NbGSTU4 proteins were expressed and purified from E. coli BL21, and resolved in a 12% SDS-polyacrylamide gel. (b) GST and GST-NbGSTU4 were UV-cross linked with r138/40A (178 nts, the 3′UTR containing the promoter for minus-strand RNA synthesis) in the presence of different concentration of glutathione (GSH) indicted on the top of each lane and resolved in a 14% SDS-polyacrylamide gel. The lane number is indicated at the bottom of each lane. (c) GST-NbGSTU4 was UV-cross linked with r138/40A in the presence of different concentrations of GSH (indicted at the top of each lane) and resolved in a 12% SDS-polyacrylamide gel. (d) GST-NbGSTU4 was incubated with BaMV r138/40A and Ba-77 (77 nts, the promoter for plus-strand RNA synthesis). The radioactive UV-crosslinked GST-NbGSTU4 protein is indicated.